• 제목/요약/키워드: catabolite repression

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Selection and Characterization of Catabolite Repression Resistant Mutant of Bacillus firmus var. alkalophilus Producing Cyclodextrin Glucanotransferase

  • Do, Eun-Ju;Shin, Hyun-Dong;Kim, Chan
    • Journal of Microbiology and Biotechnology
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    • 제3권2호
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    • pp.78-85
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    • 1993
  • In order to elucidate the mechanism which regulates the production of cyclodextrin glucanotransferase (CGTase) and to achieve overproduction of CGTase by releasing catabolite (glucose) repression, several catabolite repression resistant mutants were selected from newly screened Bacillus firmus var. alkalophilus H609, after NTG (N-methyl-N -nitro-N-nitrosoguanidine) treatment, using 2-deoxyglucose as a nonmetabolizable analog of catabolite glucose and as a selection marker. Five catabolite repression resistant mutants were selected from about 30, 000 2-deoxyglucose resistant colonies. Relative catabolite repression indices of the selected mutants were in the range of 8~80% assuming 100% for parent strain. The amount of CGTase produced by the mutant strain CR41, which was 250 units/ml, was three times larger than that produced by its parent strain. The mutation seems to have occurred in the regulatory region of CGTase gene and not in the structural region or the glucose transporting system in cell membrane. The enzymatic properties of CGTase excreted from parent and mutant strains were also compared.

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Lactobacillus sporogenes에서$\beta$-galactosidase 생합성 조절 (Regulation of $\beta$-galactosidase Biosynt hesis in Lactobacillus sporogenes)

  • 이정희;최용진
    • 한국미생물·생명공학회지
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    • 제18권6호
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    • pp.566-570
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    • 1990
  • Lactobacillus sporogenes에서 Beta-glactosidase의 생합성은 isopropyl-Beta-galactopyransoide(IPTG)나 galactose에 의해 효과적으로 유도되었으며 배양초기에는 lactose에 의해서도 훨씬 낮은 정도로 유도되었다. Glucose는 inducer exclusion이나 transient repression이 아니 catabolite repression에 의해 Beta-galactosidase의 합성을 억제시킴을 알 수 있었다. 또한, glucose에 의한 Beta-galactosidase의 catabolite repression은 cAMP나 cGMP에 의해 전혀 완하되지 않았다.

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Sisomicin발효에 대한 탄소원의 영향과 Glucose에 의한 조절효과 (Effects of Various Carbon Sources and Carbon Catabolite Regulation in Sisomicin Fermentation)

  • 안병우;이상한;신철수
    • 한국미생물·생명공학회지
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    • 제14권4호
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    • pp.293-298
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    • 1986
  • Micromonospora inyoensis 균주에 의한 sisomicin의 발효에서 항생물질의 생산성에 대한 여러 가지 탄소원의 영향을 batch culture를 이용하여 검토하였다. Starch, dextrin 및 maltose는 sisomicin의 생산에 좋은 탄소원으로 밝혀졌으나, glucose가 사용되었을 때 sisomicin 생산성은 carbon catabolite regulation에 기인하여 그게 감소되었다. 한편 sisomicin생합성에 대한 carbon catabolite regulation은 catabolite inhibition효과보다 주로 catabolite repression 효과에 좌우되었다.

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Bacillus sp. KJ16에서 Cyclodextrin Gluanotransferase와 Cyclodextrinase 생산의 Catabolite Repression

  • 김병우;권현주;이경희
    • 한국미생물·생명공학회지
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    • 제24권2호
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    • pp.137-142
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    • 1996
  • The biosynthesis and catabolite repression of cyclodextrin glucanotransferase(CGTase) and cyclodextrinase(CDase) were studied in Bacillus sp. KJI6. In accompanying to the cell growth, CGTase was synthesized during early growth phase (20h culture) and CDase was synthesized during late growth phase (60h culture). Synthesis of CGTase was rather constitutive than that of CDase in the absence or presence of carbon source. Production of CDase was strongly stimulated by amylopectin and $\gamma$-CD medium (about 6 times), but CGTase synthesis was slightly increased (about 1.3 times). Easily metabolizable carbohydrates such as D-glucose, D- fructose and D-mannose completely repressed the expression of CDase, whereas their repressive effect to CGTase synthesis was relatively negligible. By addition of 10 mM cAMP, any significant effect on the synthesis of the two enzymes was not observed. Hardly metabolizable glucose analogues such as 2-deoxy-D-glucose and 3-0-methyl-D-glucopyranose also did not show any repression on the syntheses of CGTase and CDase. This indicates that D-glucose has to be metabolized to exert its repressive effect. With these results, it seems likely that the biosynthesis of CGTase and CDase are regulated by the catabolite repression due to unknown metabolite(s) of EM pathway.

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Psedomonas sp.의 Catabolits Repression 저항성 변이주로부터 Cellulase의 생산 (Cellulase Production from the Catabolite Repression Resistant Mutant of Pseudomonas sp.)

  • 정영철;노종수;성낙계;강신권
    • 한국미생물·생명공학회지
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    • 제21권6호
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    • pp.549-555
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    • 1993
  • The production of cellulase by Pseudomonas sp. LBC505 isolated was under the strict genetic and biochemical control mechanisms such as catabolit repression and induction. These biochemical control reduced cellulase production. Thus LBC505 was mutated to increase enzyme yields. Cells growth and cellulase production were inhibited by the addition of 2-deoxy glucose (2-DG), which is presumed to function as repressor for the selection of high cellulase yielding mutant.

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혼합당에서의 Pichia stipitis의 생육 모델 (Growth model for Pichia stipitis growing on sugar mixtures)

  • 이유석;권윤중변유량
    • KSBB Journal
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    • 제7권4호
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    • pp.265-270
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    • 1992
  • 자연계에 널리 존재하는 저렴한 기질들은 대부분 여러 종류의 탄소원들을 함유한다. 이러한 혼합기질을 이용하는 효율적인 발효공정을 개발하기 위해서는 이들 기질들이 서로 어떻게 이용되는가를 알아야 하며, 사용되는 미생물의 생육과 생성물 생산을 잘 표현할 수 있는 동력학적 모델이 필요하다. P. stipitis에 의해 혼합기질에서 에탄오릉ㄹ 생산할 때 glucose는 xylose와 cellobiose 이용에 대해 ca-tabilite repression을 일으켰으며, 초기 glucose농도가 높을수록 xylose이용에서 균체의 생육속도는 감소하였으며 xylose이용시간도 길어졌다. 또한 glucose/xylose발효시 xylose이용에서 감소된 생육속도는 glucose가 xylose이용에 permenant repression을 야기시킨다는 것을 알 수 있었다. Cyclic AMP가 중개하는 catabolite repression mechanism에 기초하여 혼합기질에서 생육하는 P. stipitis의 생육모델을 발전시켰다 이 보텔식들을 이용하여 컴퓨터 simulation한 결과는 혼합기질로부터 P. stipitis에 의한 생육 및 에탄올 생성 실험결과와 비교적 잘 일치하였다.

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Carbon Catabolite Repression (CCR) of Expression of the XylanaseA Gene of Bacillus stearothermophilus No.236

  • Ha, Gyong-Sik;Choi, Il-Dong;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • 제11권1호
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    • pp.131-137
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    • 2001
  • Previous work has identified that only the catabolite responsive element A (creA; previously called cre-2) out of two potential cre sequences (cre-1: nucleotide +160 to +173 and cre-2: +173 to +186), recognized within the coding region of the xylanaseA gene (xynA) of Bacillus stearothermophilus No.236, was actually, was actually involved in the carbon catabolite repression(CCR) of xynA expression in B. subtilis. However, the level of CCR of xynA expression in the original B.stearothermophilus No.236 strain (70-fold repression). Therefore, to search for an additional cre element in the promoter region, the upstream region of the xynA gene was subcloned by chromosome walking, and as a result, another potential cre element (nucleotide -124∼-137; designated creB) was recognized in this region. The cre-like sequence revealed a high homology to the cre consensus sequence. The xylanase activity of B. subtilis MW15 bearing pWPBR14 (containing creA and creB) cultured in a medium containing xylose as the sole carbon source was about 7.7 times higher than that observed for the same culture containing glucose. B. subtilis MW15 bearing pWPBR23 (containing only creA) produced an activity about 2.4 times higher. This pattern of CCR was confirmed using derivatives of xynA::aprA fusion plasmids. Furthermore, a measurement of the amounts of the xynA transcript showed a similar pattern as that for the production of xylanase. In addition, the synthesis of xylanase in B. subtilis QB7115 [a catabolite control protein A (ccpA) mutant strain] carrying pWPBR14 was almost completely relieved from glucose repression. Together, these results lead to a conclusion that the CCR of the expression of the xynA gene is mediated by CcpA binding at creA and creB sites in B. subtilis.

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해양의 Pseudomonas sp. 로부터 분리한 alginate lyase 유전자의 promoter에 의한 대장균 내에서의 \beta-agarase 유전자의 발현과 catabolite repression의 변화 (Expression of \beta-agarase Gene and Carabolite Repression in Escherichia coli by the Promoter of Alginate Lyase Gene Isolated from Marine Pseudomonas sp.)

  • 공인수;박제현;한정현;최윤혁;이종희;진철호;이정기
    • 한국미생물·생명공학회지
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    • 제29권2호
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    • pp.72-77
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    • 2001
  • Strong promoter로 밝혀진 alginate lyase 유전자의 promoter 부위에 대한 특성을 검토하기 위해 alginate lyase 유전자의 46개 N-terminal amino acid가 포함된 promoter 부분과, 같은 균으로부터 분리한 $\beta$-agarase의 유전자를 연결시켜 agarase의 activity를 평판배지상에서 보다 쉽게 확인하는 방법으로 promoter의 활성을 측정한 결과 alginate lyase 유전자 promoter에 의해서 $\beta$-agarase 유전자의 대량발현이 유도되고 있었으며 glucose의 존재하에서 $\beta$-agarase 유전자 발현이 일어나지 않는 catabolite repression 양상을 나타내고 있다. PCR로써 alginate lyase의 46개 N-terminal amino acid 부분이 순차적으로 제거된 plasmid를 제조하여 대량발현을 조사한 결과 46개의 아미노산이 제거된 후에도 $\beta$-agarase의 활성에는 변화가 없어 46개의 N-말단이 정상적인 상태에서 발현에는 영향을 미치고 있지 않음을 확인할 수 있었다. 또한 alginate lyase 유전자의 promoter region에 존재하는 가능한 2개의 promoter consensus sequence PI, PII를 subcloning한 결과 promoter PII만이 존재할 때도 대량발현이 유도되고 있음을 확인할 수 있었으며 동시에 glucose가 존재할 때 catabolite repression이 역시 나타나고 있어 이 부분이 발현 및 glucose에 의한 regulation에 매우 중요하게 작용하는 부분이라는 것을 확인할 수 있었다.

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Catabolite Repression of the Bacillus stearothermophilus $\beta$-Xylosidase Gene (xylA) in Bacillus subtilis

  • Cho, Ssang-Goo;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • 제8권1호
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    • pp.21-27
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    • 1998
  • The xylA gene of Bacillus stearothermophilus encoding the major ${\beta}$-xylosidase was previously cloned and sequenced. In the present study we examined the regulation of the cloned xylA gene expression in Bauillus subtilis MW15 carrying the xylA::aprA fusion plasmids. The induction of the fused xylA gene expression remained uninfluenced by any of the carbon sources tested but the gene expression was repressed about 2-3 fold in the presence of glucose. Two CRE-like sequences (CRE-1: nucleotides + 124 to +136 and CRE-2: +247 to +259) were recognized within the reading frame region of the xylA gene. The deletion experiments showed that the CRE-2 sequence had a role in catabolite repression (CR) as a true CRE of the xylA gene, but the CRE-1 had no effect on CR of the xylA gene expression. Surprisingly, the deletion of the CRE- 1 sequence reduced about 2~3 fold of the expression of the xylA fused gene. The repression ratios of the xylA gene expression were estimated to be about 0.4 from the assay of subtilisin activity, and about 0.3 at the level of transcription by determining the amounts of xylA transcripts in B. subtilis. While, the level of CR of the xylA gene was assessed to be about l0-fold in previous work when the relative amounts of the xylA transcripts were measured in B. stearothermophilus.

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호알칼리성, 고온성 Bacillus cereus TA-11으로 생산된 세포내 Invertase의 생합성 조절 (Biosynthetic Regulation of Intracellular Invertase from Alkalophilic and Thermoplilic Bacillus cereus TA-11)

  • 이성훈;송정은;이종수
    • 자연과학논문집
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    • 제18권1호
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    • pp.29-38
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    • 2007
  • 호 알칼리성이며 고온성인 Bacillus cereus TA-11이 세포내로 생성하는 invertase의 생합성 조절 양상을 조사하였다. Bacillus cereus TA-11의 세포내 invertase는 10 mM sucrose의 180분 처리와 25 mM raffinose의 90분 처리에서 각각 효율적으로 유도되었다. 또한 glucose는 sucrose에 의한 invertase의 유도를 억제하였고 cAMP첨가는 catabolite repression을 감소시키지 못하였다.

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